ISSN:
1432-0878
Keywords:
Gastrin cells
;
Entero-endocrine cells
;
Rat
;
Cell isolation
;
Pylorus
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Medicine
Notes:
Summary A technique has been developed to obtain viable, isolated and enriched populations of gastrin cells (G-cells) from the rat stomach. Restricted tissue samples from a small area of the pyloric antrum known to be particularly rich in G-cells, were sequentially digested with pronase followed by mechanical agitation, to remove the epithelial cells. This technique resulted in a significant enrichment of G-cells (3–4 fold) since the surface epithelial cells and upper portions of the glands were discarded before the initial G-cell fraction was collected. These cells in suspension were then isolated from each other by gentle pipetting in a DNase containing solution and designated the crude preparation (CP). The G-cells were then purified further by separating the cells according to size by velocity sedimentation. The greatest concentration of G-cells (15–25 %) was found in the fraction containing cells with diameters of 10 to 12 μm. The effectiveness of the technique was evaluated by counting G-cells as identified by electron microscopy and immunofluorescence and assessing gastrin activity by radioimmunoassay. All three methods indicated that cell separation by gravity velocity sedimentation enriched the G-cell population 15–20 fold over their concentration in the CP. The combined techniques of selective pronase digestion followed by gravity velocity sedimentation resulted in an isolated cell preparation containing a 50–100 fold increase of G-cells over their normal distribution in the intact gastric mucosa. Since these isolated G-cells retain features indicating viability, their usefulness for in vitro studies is suggested.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00236409
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